Adeno-associated virus Rep-mediated targeting of integrase-defective retroviral vector DNA circles into human chromosome 19

► Adeno-associated virus (AAV) is capable of targeted integration in human cells. ► Integrase-defective retroviral vector (IDRV) enables a circular DNA delivery. ► A targeted integration system of IDRV DNA using the AAV integration mechanism. ► Targeted IDRV integration ameliorates the safety concer...

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Published inBiochemical and biophysical research communications Vol. 417; no. 1; pp. 78 - 83
Main Authors Huang, Shuohao, Kawabe, Yoshinori, Ito, Akira, Kamihira, Masamichi
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 06.01.2012
Subjects
60 APPLIED LIFE SCIENCES
AAVS1
Adeno-associated virus
Adeno-associated virus (AAV)
ANIMAL CELLS
CATTLE
chromosome 19
Chromosomes, Human, Pair 19 - genetics
Circular DNA
Clinical trials
Dependovirus - genetics
DISEASE VECTORS
DNA
DNA, Circular - genetics
DNA-Binding Proteins - genetics
Expression vectors
FLUORESCENCE
Gene Targeting - methods
GENE THERAPY
Genetic Vectors - genetics
Genomes
HeLa Cells
Humans
insertional mutagenesis
Integrase-defective retroviral vectors
Integrases - genetics
Integration
MUTAGENESIS
Mutagenesis, Insertional
Packaging
Plasmids
PROTEINS
Recombination, Genetic
Rep
Rep protein
Targeted integration
Transgenes
Tropism
Viral Proteins - genetics
VIRUSES
Adeno-associated virus (AAV)
RBS
Targeted integration
AAVS1
p5IEE
Integrase-defective retroviral vectors
LTR
EGFP
TRS
AAV
FBS
Rep
IDRV
Online AccessGet full text

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Abstract ► Adeno-associated virus (AAV) is capable of targeted integration in human cells. ► Integrase-defective retroviral vector (IDRV) enables a circular DNA delivery. ► A targeted integration system of IDRV DNA using the AAV integration mechanism. ► Targeted IDRV integration ameliorates the safety concerns for retroviral vectors. Retroviral vectors have been employed in clinical trials for gene therapy owing to their relative large packaging capacity, alterable cell tropism, and chromosomal integration for stable transgene expression. However, uncontrollable integrations of transgenes are likely to cause safety issues, such as insertional mutagenesis. A targeted transgene integration system for retroviral vectors, therefore, is a straightforward way to address the insertional mutagenesis issue. Adeno-associated virus (AAV) is the only known virus capable of targeted integration in human cells. In the presence of AAV Rep proteins, plasmids possessing the p5 integration efficiency element (p5IEE) can be integrated into the AAV integration site (AAVS1) in the human genome. In this report, we describe a system that can target the circular DNA derived from non-integrating retroviral vectors to the AAVS1 site by utilizing the Rep/p5IEE integration mechanism. Our results showed that after G418 selection 30% of collected clones had retroviral DNA targeted at the AAVS1 site.
AbstractList Retroviral vectors have been employed in clinical trials for gene therapy owing to their relative large packaging capacity, alterable cell tropism, and chromosomal integration for stable transgene expression. However, uncontrollable integrations of transgenes are likely to cause safety issues, such as insertional mutagenesis. A targeted transgene integration system for retroviral vectors, therefore, is a straightforward way to address the insertional mutagenesis issue. Adeno-associated virus (AAV) is the only known virus capable of targeted integration in human cells. In the presence of AAV Rep proteins, plasmids possessing the p5 integration efficiency element (p5IEE) can be integrated into the AAV integration site (AAVS1) in the human genome. In this report, we describe a system that can target the circular DNA derived from non-integrating retroviral vectors to the AAVS1 site by utilizing the Rep/p5IEE integration mechanism. Our results showed that after G418 selection 30% of collected clones had retroviral DNA targeted at the AAVS1 site.
► Adeno-associated virus (AAV) is capable of targeted integration in human cells. ► Integrase-defective retroviral vector (IDRV) enables a circular DNA delivery. ► A targeted integration system of IDRV DNA using the AAV integration mechanism. ► Targeted IDRV integration ameliorates the safety concerns for retroviral vectors. Retroviral vectors have been employed in clinical trials for gene therapy owing to their relative large packaging capacity, alterable cell tropism, and chromosomal integration for stable transgene expression. However, uncontrollable integrations of transgenes are likely to cause safety issues, such as insertional mutagenesis. A targeted transgene integration system for retroviral vectors, therefore, is a straightforward way to address the insertional mutagenesis issue. Adeno-associated virus (AAV) is the only known virus capable of targeted integration in human cells. In the presence of AAV Rep proteins, plasmids possessing the p5 integration efficiency element (p5IEE) can be integrated into the AAV integration site (AAVS1) in the human genome. In this report, we describe a system that can target the circular DNA derived from non-integrating retroviral vectors to the AAVS1 site by utilizing the Rep/p5IEE integration mechanism. Our results showed that after G418 selection 30% of collected clones had retroviral DNA targeted at the AAVS1 site.
Highlights: Black-Right-Pointing-Pointer Adeno-associated virus (AAV) is capable of targeted integration in human cells. Black-Right-Pointing-Pointer Integrase-defective retroviral vector (IDRV) enables a circular DNA delivery. Black-Right-Pointing-Pointer A targeted integration system of IDRV DNA using the AAV integration mechanism. Black-Right-Pointing-Pointer Targeted IDRV integration ameliorates the safety concerns for retroviral vectors. -- Abstract: Retroviral vectors have been employed in clinical trials for gene therapy owing to their relative large packaging capacity, alterable cell tropism, and chromosomal integration for stable transgene expression. However, uncontrollable integrations of transgenes are likely to cause safety issues, such as insertional mutagenesis. A targeted transgene integration system for retroviral vectors, therefore, is a straightforward way to address the insertional mutagenesis issue. Adeno-associated virus (AAV) is the only known virus capable of targeted integration in human cells. In the presence of AAV Rep proteins, plasmids possessing the p5 integration efficiency element (p5IEE) can be integrated into the AAV integration site (AAVS1) in the human genome. In this report, we describe a system that can target the circular DNA derived from non-integrating retroviral vectors to the AAVS1 site by utilizing the Rep/p5IEE integration mechanism. Our results showed that after G418 selection 30% of collected clones had retroviral DNA targeted at the AAVS1 site.
Author Huang, Shuohao
Kamihira, Masamichi
Kawabe, Yoshinori
Ito, Akira
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Keywords Adeno-associated virus (AAV)
RBS
Targeted integration
AAVS1
p5IEE
Integrase-defective retroviral vectors
LTR
EGFP
TRS
AAV
FBS
Rep
IDRV
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Snippet ► Adeno-associated virus (AAV) is capable of targeted integration in human cells. ► Integrase-defective retroviral vector (IDRV) enables a circular DNA...
Retroviral vectors have been employed in clinical trials for gene therapy owing to their relative large packaging capacity, alterable cell tropism, and...
Highlights: Black-Right-Pointing-Pointer Adeno-associated virus (AAV) is capable of targeted integration in human cells. Black-Right-Pointing-Pointer...
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SubjectTerms 60 APPLIED LIFE SCIENCES
AAVS1
Adeno-associated virus
Adeno-associated virus (AAV)
ANIMAL CELLS
CATTLE
chromosome 19
Chromosomes, Human, Pair 19 - genetics
Circular DNA
Clinical trials
Dependovirus - genetics
DISEASE VECTORS
DNA
DNA, Circular - genetics
DNA-Binding Proteins - genetics
Expression vectors
FLUORESCENCE
Gene Targeting - methods
GENE THERAPY
Genetic Vectors - genetics
Genomes
HeLa Cells
Humans
insertional mutagenesis
Integrase-defective retroviral vectors
Integrases - genetics
Integration
MUTAGENESIS
Mutagenesis, Insertional
Packaging
Plasmids
PROTEINS
Recombination, Genetic
Rep
Rep protein
Targeted integration
Transgenes
Tropism
Viral Proteins - genetics
VIRUSES
Title Adeno-associated virus Rep-mediated targeting of integrase-defective retroviral vector DNA circles into human chromosome 19
URI https://dx.doi.org/10.1016/j.bbrc.2011.11.059
https://www.ncbi.nlm.nih.gov/pubmed/22138242
https://search.proquest.com/docview/916520636
https://search.proquest.com/docview/926880117
https://www.osti.gov/biblio/22207619
Volume 417
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